CN109010823A - Anti- various pathogens composition and its preparation process and application, spray and its preparation process - Google Patents

Abstract

An anti-multiple pathogenic bacteria composition and its preparation process and application, a spray and its preparation process relate to the field of specific compound egg yolk antibodies. The anti-multiple pathogenic bacteria composition of the embodiment of the present invention adopts the antigen immunization that comprises tinea pedis pathogenic bacteria to obtain, and tinea pedis pathogenic bacteria comprises Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus coccus, escherichia coli, Pseudomonas aeruginosa, the anti-multiple pathogenic bacteria composition is safe, can prevent and treat tinea pedis; The preparation of tinea pedis; the spray and its preparation process of the embodiment of the present invention is to prepare the spray for preventing and treating tinea pedis by using anti-multiple pathogenic bacteria composition, which is convenient for medication, fast in absorption and remarkable in effect.

Description

Anti-multiple pathogenic bacteria composition and its preparation process and application, spray and its preparation craft

technical field

The invention relates to the field of specific composite egg yolk antibodies, and in particular to an anti-multiple pathogenic bacteria composition and its preparation process and application, spray and its preparation process.

Background technique

Tinea pedis, commonly known as athlete's foot, Hong Kong's athlete's foot, is a foot skin disease caused by pathogenic fungi and is contagious. The cause of tinea pedis is that there are no sebaceous glands that can secrete fatty acids that inhibit fungi between the soles of the feet and the toes, and the stratum corneum is rich in nutrients, sweat glands in the feet are rich, and the air circulation is poor, which is easy to form a humid and warm environment, which is very conducive to the growth of filamentous fungi. grow. Pregnant women, diabetics, or people who have used hormones and immunosuppressants for a long time are susceptible to tinea pedis because of the decline in skin resistance. The main clinical symptoms of tinea pedis include: severe itching, local abscess, skin damage, exudate, pain, easy to infect other parts of the patient, such as infection to the hands to form tinea manuum, infected fingernails (toenails) to form onychomycosis, and infection to the buttocks , Formation of jock itch at the groin, severe cases can cause secondary infections such as calf erysipelas.

At present, most antifungal drugs are used in clinical treatment of tinea pedis, such as 1%-2% econazole cream, miconazole ointment, clotrimazole ointment or other topical antifungal drugs; or griseofulvin, ketoconazole, itridine Conazole, fluconazole, terbinafine and other oral medicines. Although the use of antifungal drugs has a certain clinical effect, the side effects are relatively large, and gastrointestinal reactions are the most common. Long-term use can cause liver poisoning, leukopenia, and adverse reactions in the central nervous system.

Therefore, need to develop a kind of high security, and can prevent and treat the specific preparation of tinea pedis.

Contents of the invention

The object of the present invention is to provide an anti-multiple pathogenic bacteria composition and its preparation process, the anti-multiple pathogenic bacteria composition is safe and can prevent and treat tinea pedis.

The object of the present invention is to provide an application of the anti-multiple pathogenic bacteria composition for preparing preparations for preventing and treating tinea pedis.

The object of the present invention is to provide a spray and its preparation process. The spray for preventing and treating tinea pedis is prepared by using the anti-multiple pathogenic bacteria composition, which is convenient for medication, fast in absorption and remarkable in effect.

The present invention solves its technical problems by adopting the following technical solutions.

The present invention proposes an anti-multiple pathogenic bacteria composition, which is obtained by immunizing with antigens comprising tinea pedis pathogenic bacteria, which include Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, aureus Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa.

Further, in a preferred embodiment of the present invention, the mass ratio of Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa is 4-8:4-8: 3-5:4-8:4-8:3-5.

Further, in a preferred embodiment of the present invention, the pathogenic bacteria of tinea pedis also includes Burkholderia cepacia, and the mass ratio of Trichophyton rubrum and Burkholderia cepacia is 4-8:3-5 .

The present invention proposes a preparation process of the above-mentioned anti-multiple pathogenic bacteria composition, which comprises the following steps:

Preparation of complex antigens comprising Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa;

Carry out booster immunization to chickens with compound antigens to obtain immunized eggs of immunized chickens;

Extracting the yolk of the immunized egg to obtain dry powder of the crude extract;

The dry powder of the crude extract is separated and purified through ion exchange chromatography and gel filtration chromatography to obtain the anti-multiple pathogenic bacteria composition.

The present invention proposes an application of the anti-multiple pathogenic bacteria composition. The anti-multiple pathogenic bacteria composition is used to prepare a preparation for preventing and treating tinea pedis, and the preparation is spray, cream or lotion.

The present invention proposes a spray, which is prepared from raw materials, and the raw materials include:

Make up to 1000ml with water for injection.

Further, in a preferred embodiment of the present invention, the raw material also includes 1-5 g of Chinese medicine extract, which is obtained by extracting Chinese herbal medicines including garlic, Senecio, honeysuckle and Polygonum cuspidatum.

Further, in a preferred embodiment of the present invention, the raw material also includes 1-5 g of levofloxacin.

The present invention proposes a kind of preparation technology of above-mentioned spray, it comprises the following steps:

adding the anti-multiple pathogenic bacteria composition and suspending agent into part of the water for injection and stirring to dissolve to obtain a mixed solution;

Dissolve the borneol in ethanol, then add glycerin to mix, then add the mixed solution, and finally make up to 1000ml with water for injection to obtain the medicinal solution;

Fill 50 manual quantitative pressure spray tanks with the medicinal liquid on the filling line, each 20ml, and then fill in more than 30% of the propellant.

Further, in a preferred embodiment of the present invention, 1-5 g of Chinese medicine extract is also added when preparing the mixed solution, and the specific preparation method of the Chinese medicine extract is as follows:

Extracting the washed garlic, senecio, honeysuckle, and knotweed with water at least twice, and centrifuging the combined extracts in a high-speed centrifuge to obtain the first centrate;

The medicinal dregs after water extraction are extracted at least once with ethanol solution under reflux, and the extract is centrifuged in a high-speed centrifuge to obtain a second centrate;

The first centrifugate and the second centrifuge are combined to obtain a traditional Chinese medicine extract.

The beneficial effects of the anti-multiple pathogenic bacteria composition and its preparation process and application, spray and its preparation process of the embodiment of the present invention are: the anti-multiple pathogenic bacteria composition of the embodiment of the present invention adopts the The antigen immunization of pathogenic bacteria is obtained, and the pathogenic bacteria of tinea pedis include Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and the anti-multiple pathogenic bacteria composition is safe , can prevent and treat tinea pedis; the application of the anti-multiple pathogenic bacteria composition of the present invention is to prepare the preparation for preventing and treating tinea pedis; The spray used for preventing and treating tinea pedis is prepared from the pathogenic bacteria composition, and has the advantages of convenient medication, fast absorption and remarkable effect.

Detailed ways

In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Those who do not indicate the specific conditions in the examples are carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used were not indicated by the manufacturer, and they were all conventional products that could be purchased from the market.

The anti-multiple pathogenic bacteria composition of the embodiment of the present invention and its preparation process and application, spray and its preparation process will be described in detail below.

The embodiment of the present invention provides an anti-multiple pathogenic bacteria composition, which is obtained by immunizing with antigens including tinea pedis pathogenic bacteria, specifically preparing composite antigens by using tinea pedis pathogenic bacteria, and strengthening the chicken with the composite antigens The immunization is obtained by purifying the yolk of the immunized eggs of the obtained immunized chickens. The pathogenic bacteria of tinea pedis include Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, The mass ratio of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa is generally 4-8:4-8:3-5:4-8:4-8:3-5.

In some embodiments of the present invention, the athlete's foot pathogen can also include Burkholderia cepacia (preservation number ATCC25608), and the mass ratio of Trichophyton rubrum and Burkholderia cepacia is 4-8: 3-5. The anti-multiple pathogenic bacteria composition is the compound antibody extract against the above-mentioned tinea pedis pathogenic bacteria, and the anti-multiple pathogenic bacteria composition has a specific binding inhibitory effect on various pathogenic microorganisms that cause tinea pedis. In the case of extremely low concentration, it can also play an effective role, and it only inhibits and kills pathogenic bacteria, and it will not produce drug-resistant strains, and no toxic side effects have been found on the human body.

The embodiment of the present invention provides a preparation process of a composition against various pathogenic bacteria, which comprises the following steps:

S1. Prepare a complex antigen containing Tinea pedis pathogenic bacteria such as Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and the like.

Specifically, Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were cultured at a constant temperature of 34-36°C for 20-24 hours, and then inactivated for later use; Inactivated Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa according to mass ratio 4-8:4-8:3-5:4-8:4- 8:3-5 mixing to obtain a strain mixture; dissolve the strain mixture in a neutral, 0.08-0.1mol/L PBS solution to obtain a strain mixture solution with a concentration of 1.2-2.2mg/mL; Add Freund's adjuvant in medium volume to the mixture solution, and homogenize at high speed to obtain the composite antigen.

S2. Carrying out booster immunization to chickens with compound antigens to obtain immunized eggs of immunized chickens.

Specifically, the compound antigen is injected into the breast muscle and thigh of 20-week-old chickens, each site is injected with 0.5-0.6ml, and then boosted once every 10 days, with a total of at least 3 immunizations. The Freund's adjuvant in the compound antigen used for the first injection was incomplete Freund's adjuvant, the Freund's adjuvant in the compound antigen used after the second time was complete Freund's adjuvant, and the collection after the last injection was strengthened. Eggs laid by immunized chickens.

Carry out the chicken of booster immunity in the present embodiment is a kind of in Hailan brown chicken, Beijing red chicken, Isabeth chicken and Bailaihang laying hen; Carry out the chicken of booster immunization by following method selection: the 7th month after the first injection, mark the eggs produced by these chickens and extract the IgY therein respectively, detect the potency of the prepared IgY with ELISA method; For its titer, select the chickens that can produce IgY titer ≥ 1:128, and then use the eggs produced by these chickens to hatch chicken breeds, and wait for them to grow up to 20 weeks, as chickens for enhanced immunization.

S3. Purifying the egg yolk of the immunized egg to obtain dry powder of the crude extract.

Specifically, use an egg beater to break the immunized egg, remove the egg white, stir the egg yolk evenly, add distilled water to dilute and mix evenly according to the volume of the egg yolk 4-6 times, adjust the pH to 5.0-6.0; then cool to 2-6°C, Stand still for 12-24 hours; then centrifuge through a high-speed centrifuge for 20-30 minutes; add the separated supernatant to an ultrafilter for ultrafiltration and concentrate 10-20 times, and the mass concentration is 10%-13% Add the sodium sulfate solution into the concentrated slurry and stir thoroughly; then centrifuge through a high-speed centrifuge, take the supernatant, remove lipoproteins, and obtain the slurry; add the slurry to an ultrafiltration machine to pass through an ultrafine membrane to filter out Bacteria; freeze-dry the slurry after filtering and sterilizing with a freeze dryer to obtain dry powder of the crude extract.

S4. The dry powder of the crude extract is subjected to ion-exchange chromatography on an ion-exchange column and gel filtration chromatography on a gel-exchange column to separate and purify to obtain an anti-multiple pathogenic bacteria composition.

The embodiment of the present invention provides a kind of application of above-mentioned anti-multiple pathogenic bacteria composition, anti-multiple pathogenic bacteria composition prepares the preparation that is used for preventing and treating tinea pedis, and the dosage form of preparation can be spray, cream, lotion agent.

The embodiment of the present invention provides a spray, which is prepared from raw materials, and the raw materials include: 0.5-1.5g anti-multiple pathogenic bacteria composition; 20-25g glycerin; 3-5g borneol; 10-15g suspending agent; 5-10ml ethanol; make up to 1000ml with water for injection. The suspending agent includes at least one of microcrystalline cellulose and sodium carboxymethyl cellulose.

In some embodiments of the present invention, the raw material may also include 1-5 g of Chinese medicine extract, which is obtained by extracting Chinese herbal medicines including garlic, Senecio, honeysuckle, and Polygonum cuspidatum. Among the Chinese herbal medicines used in this example, garlic for external use has antibacterial, antiviral and antiprotozoal effects; Senecio, honeysuckle and knotweed all have the effects of clearing away heat and detoxification, and are mainly used to treat carbuncle and sore. Garlic, Senecio, Honeysuckle, and Polygonum cuspidatum are extracted to make traditional Chinese medicine extracts, which can enhance the antibacterial effect of the product and increase the antipruritic effect.

In some embodiments of the present invention, the raw material can also include 1-5g of levofloxacin, levofloxacin, as a broad-spectrum antibacterial drug, has strong antibacterial activity against most Enterobacteriaceae bacteria, and also has an antibacterial effect on Staphylococcus aureus, Adding levofloxacin can enhance the antibacterial effect of the product, thereby having a better effect on treating tinea pedis.

The spray of this embodiment uses at least one of traditional Chinese medicine extracts and levofloxacin (western medicine) to act synergistically with the anti-multiple pathogenic bacteria composition, which can further increase the antibacterial spectrum, thereby achieving the purpose of treating tinea pedis.

The embodiment of the present invention provides a preparation process of the above-mentioned spray, which comprises the following steps:

adding the anti-multiple pathogenic bacteria composition and suspending agent into part of the water for injection and stirring to dissolve to obtain a mixed solution;

Dissolve borneol in ethanol, then add glycerin and mix well, then add the mixture, and finally make up to 1000ml with water for injection;

On the filling line, transfer 50 manual quantitative pressure spray cans, each 20ml, and then fill the propellant with a mass of more than 30%.

In some embodiments of the present invention, when preparing the mixed solution, 1-5 g of Chinese medicine extract is added, and the specific preparation method of the Chinese medicine extract is as follows:

Extracting the washed garlic, senecio, honeysuckle, and knotweed with water at least twice, and centrifuging the combined extracts in a high-speed centrifuge to obtain the first centrate;

The medicinal dregs after water extraction are extracted at least once with ethanol solution under reflux, and the extract is centrifuged in a high-speed centrifuge to obtain a second centrate;

The first centrifugate and the second centrifuge are combined to obtain a traditional Chinese medicine extract.

The characteristics and performance of the present invention are described in further detail below in conjunction with the examples.

Example 1

The present embodiment provides an anti-multiple pathogenic bacteria composition, which is prepared according to the following process:

Inoculate Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa into Luria-Bertani medium respectively, and Luria-Bertani medium includes 2% of Tryptone, 1% yeast extract, 1% sodium chloride and 1.5% agar were used to culture in a tri-gas incubator for 22 hours at 35°C, and then the above strains were isolated and inoculated into broth medium In the medium, the broth medium includes 1% beef extract, 2% tryptone, 0.5% sodium chloride, and 1.5% agar in terms of mass percentages, and is cultivated in a constant temperature shaking incubator at 35°C for 26 hours. The rotating speed of the constant temperature shaking incubator is 180rpm, then inactivation is standby; With inactivated Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa by weight ratio 6: Mix 6:4:6:6:4 to obtain a strain mixture; dissolve the strain mixture in a neutral, 0.09mol/L PBS solution to prepare a strain mixture solution, and obtain a strain with a concentration of 1.8mg/mL Mixture solution; add Freund's adjuvant in an equal volume to the strain mixture solution and stir evenly to obtain a milky whole-bacteria composite antigen; put the whole-bacteria composite antigen into a high-speed homogenizer, crush and homogenize at a high speed to obtain a composite antigen .

The prepared composite antigen was injected into the pectoralis muscle and thigh of 20-week-old chickens, 0.5ml was injected into each place, and then boosted once every 10 days, and immunized four times in total. The Freund's adjuvant used for the first injection was incomplete Freund's adjuvant, and the Freund's adjuvant used for the second to fourth times was complete Freund's adjuvant. After the last injection, the chickens that had been boosted were collected Lay eggs.

Break the immunized eggs with an egg beater, remove the egg whites, stir the egg yolks evenly, add distilled water to dilute and mix evenly according to 5 times the volume of the egg yolks, adjust the pH to 6.0; then cool to 4°C, let stand for 20 hours; then centrifuge at high speed The separator was centrifuged for 25 minutes; the separated supernatant was added to an ultrafilter for ultrafiltration and concentrated 15 times, and the sodium sulfate solution with a mass concentration of 11% was added to the concentrated slurry and fully stirred; Centrifuge with a high-speed centrifuge, take the supernatant, remove lipoproteins, and obtain a slurry; put the slurry into an ultrafiltration machine to pass through an ultra-micro membrane to filter and sterilize; freeze-dry the filtered and sterilized slurry with a freeze dryer , to obtain dry powder of crude extract.

The dry powder of the crude extract is subjected to ion-exchange chromatography through an ion-exchange column and gel filtration chromatography through a gel-exchange column to separate and purify the composition to obtain an anti-multiple pathogenic bacteria composition.

Example 2

The present embodiment provides an anti-multiple pathogenic bacteria composition, which is prepared according to the following process:

Inoculate Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia into Luria-Bertani medium, Luria-Bertani medium Including 2% tryptone, 1% yeast extract, 1% sodium chloride and 1.5% agar by mass percentage, cultivated in a tri-gas incubator at 36°C for 20h, and then the above bacterial species Separate and inoculate in the broth medium, the broth medium includes 1% beef extract, 2% tryptone, 0.5% sodium chloride, 1.5% agar in terms of mass percentage, at 34 ℃ Cultivate in a constant temperature shaking incubator for 30 hours, and the rotating speed of the constant temperature shaking incubator is 200rpm, then inactivate for later use; the inactivated Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Burkholderia cepacia were mixed in a weight ratio of 8:8:3:4:4:3:4 to obtain a strain mixture; the strain mixture was dissolved in a neutral, 0.1mol/L PBS solution The strain mixture solution was prepared in , and the strain mixture solution with a concentration of 2.2 mg/mL was prepared; Freund's adjuvant was added in an equal volume to the strain mixture solution and stirred evenly to obtain a milky whole bacteria composite antigen; the whole bacteria composite Put the antigen into a high-speed homogenizer, pulverize and homogenize at a high speed, and obtain a composite antigen.

The prepared composite antigen was injected into the pectoralis muscle and thigh of 20-week-old chickens, 0.6ml was injected into each place, and then boosted once every 10 days, and immunized four times in total. The Freund's adjuvant used for the first injection was incomplete Freund's adjuvant, and the Freund's adjuvant used for the second to fourth times was complete Freund's adjuvant. After the last injection, the chickens that had been boosted were collected Lay eggs.

Break the immunized eggs with an egg beater, remove the egg whites, stir the egg yolks evenly, add distilled water to dilute and mix evenly according to 6 times the volume of the egg yolks, adjust the pH to 5.0; then cool to 6°C and let stand for 15 hours; then centrifuge at high speed The separator was centrifuged for 20 minutes; the separated supernatant was added to an ultrafilter for ultrafiltration and concentrated 10 times, and the sodium sulfate solution with a mass concentration of 10% was added to the concentrated slurry to fully stir; Centrifuge with a high-speed centrifuge, take the supernatant, remove lipoproteins, and obtain a slurry; put the slurry into an ultrafiltration machine to pass through an ultra-micro membrane to filter and sterilize; freeze-dry the filtered and sterilized slurry with a freeze dryer , to obtain dry powder of crude extract.

The dry powder of the crude extract is subjected to ion-exchange chromatography through an ion-exchange column and gel filtration chromatography through a gel-exchange column to separate and purify the composition to obtain an anti-multiple pathogenic bacteria composition.

Example 3

The present embodiment provides a spray, which is prepared according to the following preparation process:

Prepare raw materials: 1g of the anti-multiple pathogenic bacteria composition in Example 1; 20g of glycerin; 4g of borneol; 10g of microcrystalline cellulose; 6ml of ethanol; water for injection.

Add the anti-multiple pathogenic bacteria composition and microcrystalline cellulose into 500ml of water for injection and stir to dissolve to obtain a mixed solution;

Dissolve borneol in ethanol, then add glycerin and mix well, then add the mixture, and finally make up to 1000ml with water for injection;

On the filling line, transfer 50 manual quantitative pressure spray cans, each 20ml, and then fill more than 30% of the propellant.

Example 4

The present embodiment provides a spray, which is prepared according to the following preparation process:

1g anti-multiple pathogenic bacteria composition in embodiment 1; 20g glycerin; 3g borneol; 10g sodium carboxymethylcellulose; 5ml ethanol; 2g Chinese medicine extract (obtained by garlic, Senecio, honeysuckle, Polygonum cuspidatum extraction); Water for Injection.

Add the anti-multiple pathogenic bacteria composition, traditional Chinese medicine extract, and sodium carboxymethylcellulose into 600ml of water for injection and stir to dissolve to obtain a mixed solution;

Dissolve borneol in ethanol, then add glycerin and mix well, then add the mixture, and finally make up to 1000ml with water for injection;

On the filling line, transfer 50 manual quantitative pressure spray cans, each 20ml, and then fill more than 30% of the propellant.

Example 5

The present embodiment provides a spray, which is prepared according to the following preparation process:

1g anti-multiple pathogenic bacteria composition in Example 2; 20g glycerin; 3g borneol; 10g microcrystalline cellulose; 5ml ethanol; 2g levofloxacin; water for injection.

Add the anti-multiple pathogenic bacteria composition, levofloxacin, and sodium carboxymethylcellulose into 600ml of water for injection and stir to dissolve to obtain a mixed solution;

Dissolve borneol in ethanol, then add glycerin and mix well, then add the mixture, and finally make up to 1000ml with water for injection;

On the filling line, transfer 50 manual quantitative pressure spray cans, each 20ml, and then fill more than 30% of the propellant.

Example 6

The present embodiment provides a spray, which is prepared according to the following preparation process:

1g anti-multiple pathogenic bacteria composition in embodiment 2; 22g glycerin; 4g borneol; 12g sodium carboxymethylcellulose; 6ml ethanol; 1g Chinese medicine extract (obtained by extracting garlic, Senecio, honeysuckle, Polygonum cuspidatum); 1g levofloxacin; water for injection.

Add the anti-multiple pathogenic bacteria composition, traditional Chinese medicine extract, levofloxacin, and sodium carboxymethylcellulose into 600ml of water for injection and stir to dissolve to obtain a mixed solution;

Dissolve borneol in ethanol, then add glycerin and mix well, then add the mixture, and finally make up to 1000ml with water for injection;

On the filling line, transfer 50 manual quantitative pressure spray cans, each 20ml, and then fill more than 30% of the propellant.

The performance and therapeutic effects of the sprays of the embodiments of the present invention are investigated through tests below.

1. Skin toxicity test: Take 30 healthy adult rabbits and divide them into 5 groups. Each group includes 3 intact skins and 3 damaged skins. One group is a blank control group without any treatment; the other 4 groups are test groups. The sprays of Examples 3-6 were used to spray 4 groups of rabbits in the test group respectively, and each rabbit was sprayed 3 times a day, 3 sprays each time, for 7 days. The respiration, heart rate and limb activity of all rabbits were measured.

The results showed that there was no statistically significant difference between the test group and the blank control group, and no systemic toxic and side effects were found.

2. Clinical trials

Select 100 cases of tinea pedis patients, symptoms: interdigital erosion, blisters, stench, itching, patients age range 30-60 years old, 60 cases of males, 40 cases of females. The curative effect standard is as follows:

Cure: the feet are erosive and dry, no itching, no blisters, no desquamation;

Effective: less blisters, less itching, slight desquamation;

Ineffective: No obvious effect, increased blisters, local itching, and desquamation.

100 routine tinea pedis patients were divided into 5 groups, which were respectively control group and test group 1-4. The control group was not given any treatment, and test group 1-4 was correspondingly treated with the spray of embodiment 3-6. Therapeutic method is: adopt spray to spray medicine to the foot of tinea pedis patient, take medicine 3 times every day, press 3 sprays every time medicine. The treatment effects of each group were counted after two weeks of medication, as shown in Table 1.

Table 1 Tinea pedis treatment effect

It can be seen from the above table that the effective rate of treating tinea pedis of the spray prepared by the anti-multiple pathogenic bacteria composition of the embodiment of the present invention is more than 90%, and the curative effect is remarkable.

In summary, the anti-multiple pathogenic bacteria compositions of the embodiments of the present invention are safe and can prevent and treat tinea pedis; The preparation of tinea pedis; the spray and its preparation process of the embodiment of the present invention is to prepare the spray for preventing and treating tinea pedis by using anti-multiple pathogenic bacteria composition, which is convenient for medication, fast in absorption and remarkable in effect.

The embodiments described above are some, not all, embodiments of the present invention. The detailed description of the embodiments of the invention is not intended to limit the scope of the claimed invention but to represent only selected embodiments of the invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

Claims (10)

1. an anti-multiple pathogenic bacteria composition is characterized in that, it adopts the antigen immunization that comprises tinea pedis pathogenic bacteria to obtain, and described tinea pedis pathogenic bacteria comprises Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa. 2. anti-multiple pathogenic bacteria compositions according to claim 1, is characterized in that, described Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa The mass ratios are 4-8:4-8:3-5:4-8:4-8:3-5, respectively. 3. anti-multiple pathogenic bacteria compositions according to claim 1, is characterized in that, described tinea pedis pathogenic bacteria also comprises Burkholderia cepacia, described trichophyton rubrum and described primary cepacia The mass ratio of Kholderia is 4-8:3-5. 4. a preparation process of anti-multiple pathogenic bacteria composition as claimed in claim 1, is characterized in that, it comprises the following steps: Preparation of complex antigens comprising Trichophyton rubrum, Epidermophyton flocculus, Candida albicans, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa; Using the composite antigen to boost immunization of chickens to obtain immunized eggs of immunized chickens; Extracting the egg yolk of the immunized egg to obtain dry powder of the crude extract; The dry powder of the crude extract is separated and purified by ion exchange chromatography and gel filtration chromatography to obtain the anti-multiple pathogenic bacteria composition. 5. an application of anti-multiple pathogenic bacteria composition as claimed in claim 1, is characterized in that, described anti-multiple pathogenic bacteria composition prepares the preparation that is used for preventing and treating tinea pedis, and described preparation Available as a spray, cream, or lotion. 6. A spray, characterized in that it is prepared from raw materials, and the raw materials include: Make up to 1000ml with water for injection. 7 . The spray according to claim 6 , wherein the raw material further comprises 1-5 g of Chinese medicine extract, and the Chinese medicine extract is obtained by extracting Chinese herbal medicines including garlic, Senecio, honeysuckle, and Polygonum cuspidatum. 8. The spray according to claim 6, characterized in that, the raw material also comprises 1-5g levofloxacin. 9. a preparation process of spray as claimed in claim 6, is characterized in that, it comprises the following steps: adding the anti-multiple pathogenic bacteria composition and suspending agent into part of the water for injection and stirring to dissolve to obtain a mixed solution; Dissolve the borneol in ethanol, add glycerin and mix evenly, then add the mixed solution, and finally make up to 1000ml with water for injection to obtain the medicinal solution; The medicinal liquid is filled into 50 manual quantitative pressure spray tanks on the filling line, each 20ml, and then filled with more than 30% propellant. 10. the preparation process of spray according to claim 9 is characterized in that, also add 1-5g Chinese medicine extract when preparing mixed solution, the concrete preparation method of described Chinese medicine extract is: extracting the washed garlic, senecio, honeysuckle, and knotweed with water at least twice, and centrifuging the combined extracts in a high-speed centrifuge to obtain the first centrate; The medicinal dregs after water extraction are extracted at least once with ethanol solution under reflux, and the extract is centrifuged in a high-speed centrifuge to obtain a second centrate; Combine the first centrifugate and the second centrifuge to obtain the Chinese medicine extract.